Ultrastructure of cel organelles by scanning electron microscopy of thick sections surface-etched by an oxygen plasma

Kidney tissue double fixed in glutaraldehyde and osmium tetroxide and embedded in epoxy resin by standard techniques used for transmission electron microscopy was cut into section 1 micron or more thick and surface-etched by an oxygen plasma. Etching caused ash residues (possibly composed partly or organo-metallic complexes) of membranes and other etch resistant cell components to emerge as recognizable structures projecting upward from the surrounding embedment which was combusted and removed as volatile products. using the secondary electron mode for image formation, structural features of cells which could be imaged with clarity with the scanning electron microscopy included: profiles of peripheral and in-folded plasma membranes, the nuclear envelope and profiles of cut mitochondrial matrix granules, cristae and the outer limiting membranes. Resolution was better than that obtainable from most other methods of specimen preparation currently being used in scanning electron microscopy for viewing the internal structures of cells or organelles in bulk samples of tissue.